Purification and functional interactions of GRASP55 with Rab2

Abstract

GRASP55 is a member of the GRASP family of peripheral membrane proteins thought to contribute to the organization of the Golgi apparatus. GRASPs are typically found in complex with coiled-coil protein, termed golgins, which interact with specific GTPases of the RAB family. Here I will describe the purification of native GRASP55 complexes from Golgi membranes, and the comparison with the related GRASP65 complex, standard methods for the analysis of interactions between GRASPs, golgins, and Rab GTPases using yeast two-hybrid analysis and protein biochemistry with native and recombinant proteins are also described.